• Journal of Microbiology and Biotechnology
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• 제23권5호
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• pp.689-698
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• 2013

The production and characterization of an active recombinant N-acetylgalactosamine-6-sulfate sulfatase (GALNS) in Escherichia coli BL21(DE3) has been previously reported. In this study, the effect of the signal peptide (SP), inducer concentration, process scale, and operational mode (batch and semi-continuous) on GALNS production were evaluated. When native SP was presented, higher enzyme activity levels were observed in both soluble and inclusion bodies fractions, and its removal had a significant impact on enzyme activation. At shake scale, the optimal IPTG concentrations were 0.5 and 1.5 mM for the strains with and without SP, respectively, whereas at bench scale, the highest enzyme activities were observed with 1.5 mM IPTG for both strains. Noteworthy, enzyme activity in the culture media was only detected when SP was presented and the culture was carried out under semi-continuous mode. We showed for the first time that the mechanism that in prokaryotes recognizes the SP to mediate sulfatase activation can also recognize a eukaryotic SP, favoring the activation of the enzyme, and could also favor the secretion of the recombinant protein. These results offer significant information for scaling-up the production of human sulfatases in E. coli.

• Mycobiology
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• 제40권4호
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• pp.236-243
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• 2012

Pleurotus nebrodensis is an edible and commercially available mushroom in Korea. This study was conducted in order to evaluate the anticancer and immunopotentiating activities of crude polysaccharides, extracted in methanol, neutral saline, and hot water (hereafter referred to as Fr. MeOH, Fr. NaCl, and Fr. HW, respectively) from the fruiting bodies of P. nebrodensis. ${\beta}$-Glucan and protein contents in Fr. MeOH, Fr. NaCl, and Fr. HW extracts of P. nebrodensis ranged from 23.79~36.63 g/100 g and 4.45~6.12 g/100 g, respectively. Crude polysaccharides were not cytotoxic against sarcoma 180, HT-29, NIH3T3, and RAW 264.7 cell lines at a range of $10{\sim}2,000{\mu}g/mL$. Intraperitoneal injection with crude polysaccharides resulted in a life prolongation effect of 11.76~27.06% in mice previously inoculated with sarcoma 180. Treatment with Fr. NaCl resulted in an increase in the numbers of spleen cells by 1.49 fold at the concentration of $50{\mu}g/mL$, compared with control. Fr. HW improved the immuno-potentiating activity of B lymphocytes through an increase in alkaline phosphatase activity by 1.65 fold, compared with control at $200{\mu}g/mL$. Maximum production of nitric oxide ($14.3{\mu}M$) was recorded in the Fr. NaCl fraction at $200{\mu}g/mL$. Production of tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$), interleukin-$1{\beta}$ (IL-$1{\beta}$), and interleukin-6 (IL-6) was significantly higher, compared to control, and IL-6 production was highest, in contrast to TNF-${\alpha}$, IL-$1{\beta}$, and positive control, concanavalin at the tested concentration of the various fractions. Results of the current study suggest that polysaccharides extracted from P. nebrodensis have a strong anticancer effect and may be useful as an ingredient of biopharmaceutical products for treatment of cancer.

• Preventive Nutrition and Food Science
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• 제19권2호
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• pp.115-123
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• 2014

In the current study, wheat flour was mixed with high quality cassava flour (HQCF) in several ratios: 90:10, 80:20, 70:30, and 60:40, and used to prepare 10%, 20%, 30%, and 40% National Root Crops Research Institute (NRCRI) cassava bread, respectively. 100% wheat bread was prepared as a control (100% wheat bread). Five bread samples were prepared per group. Antioxidant assays [i.e., 2,2-diphenyl-1-picrylhydrazyl radical (DPPH) scavenging assay, reducing power assay] revealed that the bread samples had considerable antioxidant capacities. Substitution of wheat flour with HQCF at various concentrations resulted in dose dependent decreases in the mineral and protein contents of the resulting bread samples. The crude fiber content of the bread samples was minimal, while the carbohydrate content of the bread samples ranged from 43.86% to 48.64%. A 20% substitution of wheat flour with HQCF yielded bread samples with a general acceptability that was comparable to that of 100% wheat bread. The mean bacteria counts of the bread samples ranged from $2.0{\times}10^3CFU/mL$ to $1.4{\times}10^4CFU/mL$, while the fungal counts ranged from 0 CFU/mL to $3{\times}10^3CFU/mL$. There was a positive correlation between the DPPH antioxidant activities and the reducing powers of the bread samples ($R^2=0.871$) and a positive correlation between the DPPH antioxidant activities and the flavonoid contents of the bread samples ($R^2=0.487$). The higher microbial load of the NRCRI cassava bread samples indicates that these bread samples may have a shorter shelf life than the 100% wheat bread. The significant positive correlation between total flavonoid content and reducing power ($R^2=0.750$) suggests that the flavonoids present in the lipophilic fractions of the bread samples could be responsible for the reductive capacities of the bread samples.

• 대한의생명과학회지
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• 제14권3호
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• pp.147-155
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• 2008

In previous works, we found that solvent extract of Opuntia humifusa Raf., a member of the lactaceae family, displayed potent anti-oxidative and anti-inflammatory activities. Thus, all solvent fractions, except for the water layer, showed potent scavenging effects. According to activity-guided fractionation, one of active radical scavenging principles in the ethyl acetate fraction was found to be taxifolin. In this study, we investigated whether taxifolin showed anti-oxidative activity. In addition, taxifolin modulated nitric oxide (NO) release and the expression of pro-inflammatory cytokine mRNA such as interleukin-$1{\beta}$ (IL-$1{\beta}$), IL-6, granulocyte-macrophage colony-stimulating factor (GM-CSF), and TNF-${\alpha}$. Taxifolin showed potent anti-oxidant activity with the $IC_{50}\;of\;8.5{\pm}1.4\;and\;9.3{\pm}1.0{\mu}M$ using xanthine/xanthine oxidase (XO) assay and 2,2-Diphenyl-lpicrylhydrazyl radical (DPPH) assay, respectively. We next determined the role of taxifolin on the immunomodulating activity using murine macrophage cell line RAW264.7 cells. Taxifolin dose-dependently inhibited NO production in lipopolysaccharide (LPS)-activated RAW264.7. It also significantly blocked the expression of inducible NO synthase (iNOS) mRNA in the LPS-stimulated RAW264.7 cells. In addition, taxifolin potently suppressed the expression of IL-$1{\beta}$, IL-6 and GM-CSF mRNA in LPS-activated RAW264.7 cells, but not that of TNF-${\alpha}$ Moreover, taxifolin significantly inhibited the transcriptional activity of nuclear factor-${\kappa}B$ (NF-${\kappa}B$) and activator protein -1 (AP-1). These results suggest that taxifolin may downregulate inflammatory iNOS, IL-$1{\beta}$, IL-6 and GM-CSF gene expressions through inhibition of NF-K and AP-1 activation in LPS-stimulated RAW264.7 cells.

• 생명과학회지
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• 제13권5호
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• pp.668-674
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• 2003

홍어를 발효하여 발효기간에 따른 항고혈압 효과를 조사하고 그에 따른 항고혈압성 물질을 분리하기 위해 GPC system을 사용하여 항고혈압 물질을 조 분리하였다. 항고혈압성 물질을 농축하기 위해 홍어 내장 및 가식부 열수 추출물을 대량 포집 하였다. 그리고 그에 따른 ACE 억제효과를 검색하였다. 즉, 홍어 가식 부의 ACE 저해 작용은 2% 첨가시 29%로, 다소 낮으나 상시 섭취될 수 있는 식품이란 측면에서 볼 때 그 유용성이 기대된다고 할 수 있으며 홍어 내장 열수 추출물의 ACE 저해 효과는 시료 2% 첨가의 경우 발효 0일째에 71.0%로 가장 높게 나타났다. 발효가 진행되면서 ACE 저해 효과는 감소하는 것으로 나타났다. 이러한 ACE 억제효과를 나타내는 성분을 추정하기 위하여 홍어 내장 열수 추출물의 일반성분과 원소분석을 실시한 결과 일반성분 중 순단백질이 58.7%로 가장 높게 나타났으며, 원소분석 결과 C, H, O, N의 성분비가 당류라기 보다는 peptide계인 것으로 나타났다. 또한, 질소화합물의 분석결과 ACE를 억제하는 기능성 peptide의 성분인 Tyr, Phe, Val, His 등이 가식부 보다 많아서 ACE를 억제하는 peptide를 함유하리라 예상되었다. ACE억제 물질을 조 분리하기 위하여 Sephadex G-25 column chromatography에 의해 분자량별로 분획한 결과, 분획물 들의 ACE 저해 효과는 분획물 B(111-160)의 농도 0.2%에서 67.8%로 가장 높은 ACE저해 효과를 나타내었다. 이상의 결과로 미루어 보아 홍어 내장 열수 추출물의 ACE 저해인자는 가열에 대하여 안정한 비교적 저분자의 peptide와 같은 물질이라고 추정할 수 있었다.

• 한국축산식품학회지
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• 제31권4호
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• pp.563-569
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• 2011

일상 생활에서 섭취하는 가열한 생선, 육류 등의 고단백질 식품에서는 돌연변이원, 발암가능성 물질인 heterocyclic amine류가 생성된다. 안전성 측면을 고려하고자 백련 잎을 이용한 마리네이드 소스를 첨가하여 쇠고기의 HCAs의 형성억제효과와 돌연변이원성 억제효과를 알아보았다. Ames assay 결과 백련 잎 butanol fraction을 2.0 g 첨가하여 가열한 쇠고기 스테이크에서 61.5%로 높은 돌연변이원성 억제효과를 나타내었지만 용매 별 fraction 첨가군 사이에서는 유의적인 차이를 나타내지 않았다. 가열 전 쇠고기 마리네이드 소스에 식용이 가능한 백련잎 water fraction(2.0, 4.0, 8.0 g)을 첨가한 후 $190^{\circ}C$에서 가열한 결과 MeIQx(2-amino-3,8 dimethylimidazo[4,5-f]-quinoxalin)는 30.9-63.5%, PhIP(2-amino-1-methyl-6-pheny-limidazo[4,5-b]-pyridine)는 31.6-60.7% 억제되었다. $225^{\circ}C$에서의 가열된 쇠고기에서는 MeIQx가 38.1-65.3%, DiMeIQx (2-amino-3,4,8 trimethylimidazo[4,5-f]-quinoxaline)는 36.8-73.9%, PhIP는 33.9- 67.6%의 억제효과를 보였다. 따라서 백련 잎 추출물은 heterocyclic amine류 형성과 돌연변이 원성을 억제하는 효과가 있는 것으로 나타나 고단백질 식품 섭취시 안전성 측면에서 긍정적인 영향을 미칠 것으로 판단된다.

• Asian-Australasian Journal of Animal Sciences
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• 제13권12호
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• pp.1674-1680
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• 2000

A study was conducted to compare the feeding value of urea treated and untreated mustard straw (MS) for sheep. Treated MS was prepared by adding urea-N at 1.84% and followed by packing in a pit silo for 21 days. Two groups of six empty Avikaline ewes were fed untreated (UTMS) and treated (TMS) mustard straw along with 200 g concentrate per head daily for 90 days. Untreated MS had 0.41% N and the urea treatment increased its N value to 1.58 %. The cell wall constituents were decreased in the TMS except for cellulose which remained unaffected. Dry matter intake of TMS was consistently higher than that of UTMS. Digestibility of DM, OM and fibre fractions of MS improved by the urea treatment. Ewes in both groups were in positive N balance while % N retention was lower in UTMS (26.30%) than in TMS (52.14%). The TMS fed group on average consumed 30.2 g DM, 2.9 g digestible crude protein and $0.2MJ\;DE\;per\;kg\;BW\;day^{-1}$ and maintained their weight whereas, the UTMS fed ewes lost weight. The VFA concentration in rumen liquor was higher in TMS than in UTMS. Total-N, ammonia-N and TCA-precipitable-N were also higher in TMS fed ewes. Blood glucose concentrations in the two groups were similar at initiation of the study. However the glucose concentration of UTMS fed group was significantly (p<0.01) lower than those fed UTMS at the termination of the study. Urea-N concentration was also higher in TMS fed group after 90 days of feeding period. It is concluded that urea treatment of MS improved N value of MS from 0.41% to 1.58% along with sizable improvement in nutritive value and in conjunction with 200 g concentrate, TMS can serve as maintenance ration for sheep. ($ME_{lakt}/ME_{m}=1.46$).

• Asian-Australasian Journal of Animal Sciences
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• 제6권2호
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• pp.291-300
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• 1993

The agronomic, morphologic and nutritive measurements were determined for ten varieties of the early-, medium- and late- maturing rice from five types of soil in south of China. The results are shown that (1) The higher contents of neutral detergent fibre (NDF), cellulose (CEL), hemicellulose (HC) and lignin (LIG), but lower crude protein (CP) and neutral detergent solubles (NDS) contents were noted for the whole plant of rice straw during maturation; (2) As far as the feed nutritive value, segments (S) is highest, then leaf blades (LB), leaf sheaths (LS) lowest. However, LB and LS are constituted about 75% of whole plant, the nutritive value of rice straw is depended upon the nutritive quality of LB and LS; (3) The dry matter disappearances (DMD) values of different spots of rice straw are different, the eary-maturing highest, then the medium; the late- lowest; (4) The DMD value of different fractions is different, S highest, then LB, LS lowest; (5) The different retention time in rumen, the DMD value of rice straw is different. As time following, the DMD value increased gradually, during 48-72 h, the DMD value achieves close to highest; (6) The grain yield (r = -0.91), plant height (r = -0.87) and full-filling grain percent (r = -0.75) are correlated negatively with DMD value, but the leaf/stem (r = 0.59) and the proportion of stem (r = 0.58) are correlated positively with DMD value. The relations between chemical compositions and DMD value are: Early-: DMD = 7.372 + 0.055 DM - 0.532 CP - 2.487 NDF + 1.143 ADF + 0.214 CEL + 1.456 HC + 0.718 LIG (r = 0.61). Medium-: DMD = 333.927 + 2.026 DM - 0.224 CP - 4.602 NDF + 4.524 ADF + 0.149 CEL + 2.923 HC + 0.035 LIG (r = 0.79). Late-: DMD = 133.284 + 0.282 DM - 3.455 CP - 22.185 NDF + 24.267 ADF + 0.316 CEL - 23.288 HC + 0.945 LIG (r = 0.79). Therefore, it is possible to predict the nutritive value of rice straw on the basis of the agronomic, morphologic measurements and chemical compositions and the relationship with DMD value.

• 자연과학논문집
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• 제9권1호
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• pp.19-24
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• 1997

PCR 방법을 이용하여 K11 RNA 중합효소를 coding하는 Klebsiella phage gene 1을 cloning 하였고 lac 전사촉진제 조절 하에 발현시켰다. K11 RNA 중합효소는 DAEA-sephacel과 Affigel blue column chromatographies를 사용하는 상용 방법으로 분리하였다. DAEA-sephacel의 0.2-0.3 M $NH_4Cl$ 분획에서 K11 RNA 중합효소의 활성을 보였고, 다음 단계의 Affigel blue column에서 SDS-polyacryl amide gel 상의 단일 band로 분리되었다. K11 RNA 중합효소는 T7 그룹 phage RNA 중합효소로 다른 T7 그룹phage RNA 중합효소와 많은 상동성을 보인다. (대장균 phage T7, T3과 Salmonella tyhimurium phage SP6 RNA 중합효소). 이미 우리는 T7과 SP6 전사촉진제 변이체를 제조한 바 있고 T7과 SP6 RNA 중합효소의 전사촉진제 특이성을 연구한 바 있다 (이상수와 강창원, 1993). K11 RNA 중합효소의 전사촉진제 특이성을 알아보기 위해 SP6 전사촉진제 변이체를 사용하여 in vitro K11 RNA 중합효소의 활성을 측정하였다. 이 변이체 중 K11 전사촉진제와 가장 유사한 것이 가장 높은 K11 RNA 중합효소 활성을 보였다.

• 대한한방내과학회지
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• 제31권1호
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• pp.153-165
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• 2010

Objectives : The aims of this study were to investigate the cytoprotective, antioxidative and inflammation genes inhibitory effects of Patriniae Radix on the mouse LLC-$PK_1$ cells (renal epithelial cells). Methods : The cytoprotective effect of Patriniae Radix was evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The antioxidative effect was measured in terms of generation amount of superoxide anion radical (${\cdot}{O_2}^-$) by 2',7'-dichlorodihydrofluorescein diacetate (DCFDA), nitric oxide (NO) by 4,5-diaminofluorescein (DAF-2), peroxynitrite ($ONOO^-$) by dihyldrorhodamine 123 (DHR 123) and prostaglandin $E_2$ ($PGE_2$) by $PGE_2$ immunoassay on $H_2O_2$-treated LLC-$PK_1$ cells. For measuring of inflammation genes inhibitory effects, western blot was performed to detect IKK-$\alpha$, phospho-$I{\kappa}B-\alpha$, NF-${\kappa}B$ (p50, p65), COX-2, iNOS, IL-$1{\beta}$ and VCAM-1 protein level in cytosol fractions from LLC-$PK_1$ cells. Results : Patriniae Radix extract reduced the $H_2O_2$-induced cell death and inhibited the amount of $H_2O_2$-induced ${\cdot}{O_2}^-$, NO, $ONOO^-$, $PGE_2$ generation dose-dependently on the mouse LLC-$PK_1$ cells in vitro. Also Patriniae Radix extract inhibited the expression of IKK-$\alpha$, phospho-$I{\kappa}B-\alpha$, COX-2, iNOS, IL-$1\beta$ and VCAM-1 genes dose-dependently by means of decreasing activation of NF-${\kappa}B$. Conclusions : According to above results, it was identified that Patriniae Radix had the cytoprotective, antioxidative and inflammation genes inhibitory effects. So it was suggested that Patriniae Radix would be effective to the treatment for the inflammatory process and inflammation-related diseases.